Study On The Inhibitory Effect And Mechanism Of Sanguinarine On Cisplatin Resistant Ovarian Cancer

ObjectivesTo explore the effect and mechanism of sanguinarine on cisplatin resistant ovarian cancer cells and transplanted tumor in nude mice,and to provide experimental and theoretical basis for Sanguinarine combined with cisplatin in the treatment of cisplatin resistant ovarian cancerMethods:SKOV3,SKOV3/DDP,A2780 and A2780/DDP ovarian cancer cells were cultured.Each cell line was divided into control group,cisplatin group,sanguinarine group and sanguinarine combined cisplatin group,after 4 hours,10 ul of Phosphate Buffer Saline(PBS),cisplatin(2.24umol/l),sanguinarine(2.5ug/l)and sanguinarine combined cisplatin(2.24umol/l+2.5ug/l)were added to each hole of each group.①MTT method was used to detect the cell proliferation of each group at 0h,6h,12h,24h,48h and 72h,Study on the inhibitory effect of sanguinarine on ovarian cancer cells and cisplatin resistant ovarian cancer cells;②BALB/c nude mice transplanted tumor model was established by SKOV3 and SKOV3/DDP cells,and was randomly divided into 4 groups(n=3):control group,cisplatin group,sanguinarine group,sanguinarine combined cisplatin group,PBS,cisplatin(1.6mg/ml),sanguinarine(2mg/ml),sanguinarine(4mg/ml)+cisplatin(3.2mg/ml)were injected subcutaneous near the tumor with 50 ul/mouse respectively;Once every 3 days,5 times.During the experiment,the tumor size and mouse weight were measured every two days,and the growth curve of mouse weight and tumor volume were drawn to observe the anti-tumor effect;after the natural death of the mice or 5 times of drug use,kill the mice and take out the tumor to measure the weight.③Remove the tumor tissue,the expression of AREG,EGFR,pJNK,cJUN and Ki67 in the tumor tissues was detected by immunohistochemistry.Results:1.In vitro MTT test showed that the growth of ovarian cancer sensitive cell lines SKOV3,A2780 and cisplatin resistant cell lines SKOV3/DDP,A2780/DDP in the cisplatin,sanguinarine,sanguinarine combined with cisplatin groups were significantly inhibited compared with the control group and the inhibitory effect were the most obvious in sanguinarine combined with cisplatin group;Compared with cisplatin group,the inhibitory effect of sanguinarine group on ovarian cancer sensitive cells SKOV3 and A2780 was similar,there was no statistical significance(P>0.05).The inhibitory effect of sanguinarine combined with cisplatin group was significantly stronger than that of cisplatin group(P<0.05);while the inhibitory effect on ovarian cancer resistant cells SKOV3/DDP and A2780/DDP were significantly stronger in sanguinarine,sanguinarine combined with cisplatin group than that in cisplatin group,the effect in the combined group was more obvious,and the difference was statistically significant(P<0.05).2.The experimental results of transplanted tumor in mice showed that the growth of SKOV3 and cisplatin resistant SKOV3/DDP transplanted tumors cisplatin,sanguinarine,sanguinarine combined with cisplatin groups were significantly inhibited compared with the control group,the combination of sanguinarine and cisplatin showed the most significant inhibitory effect;Compared with cisplatin group,the inhibitory effect of sanguinarine on ovarian cancer sensitive SKOV3 transplanted tumor was similar,there was no significant difference(P>0.05),the inhibitory effect of sanguinarine combined with cisplatin group was significantly stronger than that of cisplatin group(P<0.05);However,the inhibitory effect on ovarian cancer resistant SKOV3/DDP transplanted tumor was significantly stronger in sanguinarine,sanguinarine combined with cisplatin group than that of cisplatin group,the effect of the combined group was more obvious,and the difference was statistically significant(P<0.05).There was no significant change in the weight of mice in each experimental group(P>0.05).3.Immunohistochemical results showed that the expression of AREG,EGFR,cJUN,pJNK and Ki67 protein in SKOV3 and SKOV3/DDP were significantly down regulated in the sanguinarine,sanguinarine combined with cisplatin group compared with the control group(P<0.05);The protein levels of AREG,EGFR,pJNK,cJUN,Ki67 in SKOV3 transplanted tumors and EGFR,cJUN,Ki67 in SKOV3/DDP transplanted tumors were significantly decreased in cisplatin group(P<0.05),while the expression levels of AREG and pJNK in SKOV3/DDP transplanted tumors were not significantly decreased compared with the control group,The difference was not statistically significant(P>0.05);Compared with cisplatin group,the protein expression levels of AREG,EGFR,cJUN,pJNK and Ki67 were similar in the sensitive ovarian cancer cell transplantation tumor tissues of sanguine group,and there was no significant difference(P>0.05),The protein expression of AREG,EGFR,cJUN,pJNK and Ki67 in the transplanted tumor tissue of resistant ovarian cancer cells was significantly decreased in the group of sanguinarine,sanguinarine combined with cisplatin(P<0.05),while that in the group of sanguinarine combined with cisplatin was significantly decreased(P<0.05).Conclusion:1.sanguinarine can significantly inhibit the growth of cisplatin resistant ovarian cancer cells in vivo and vitro experiment,and enhance the inhibitory effect of cisplatin on drug-resistant ovarian cancer.2.Sanguinarine can inhibit the phosphorylation of JNK,reduce the expression of cJUN,block the EGFR/ErbB2 signaling pathway.promote the sensitivity of cisplatin to chemotherapy,and improve the anti-tumor effect of cisplatin.